翼内肌肌浆网钙腺苷三磷酸酶的卡因差异作用。

Differential Effect of Articaine on Sarcoendoplasmic Reticulum Calcium Adenosine Triphosphatase of Medial Pterygoid Muscle.

出版信息

J Oral Facial Pain Headache. 2017;31(4):e21-e28. doi: 10.11607/ofph.1835.

Abstract

AIMS

To determine the effect of articaine on sarcoendoplasmic reticulum calcium adenosine triphosphatase (SERCA) isoforms of the medial pterygoid muscle.

METHODS

Native SERCA from the medial pterygoid muscles of 24 rabbits was isolated by ultracentrifugation, and its isoforms were purified by chromatography and assessed by enzyme-linked immunosorbent assay (ELISA). SERCA activity and calcium transport capability were determined by using colorimetric and radioisotopic methods. The mean ± standard deviation (SD) half maximal inhibitory concentration (IC50) of articaine was determined for each isoform, and these values were compared by using analysis of variance (ANOVA) (P < .05).

RESULTS

The native SERCA preparation consisted of 34% SERCA1a, 53% SERCA2a, 10% SERCA2b, and 3% combined SERCA3 and SERCA1b. Articaine caused inhibition of activity and calcium uptake in the native SERCA preparation and in each of the purified isoforms. The IC50 (mM) values for enzymatic activity were: SERCA1a 22.0 ± 2.3 > SERCA2a 16.4 ± 2.4 > SERCA2b 11.3 ± 1.9, and 15.1 ± 2.1 for native SERCA. For calcium transport, IC50 values were: SERCA1a 31.1 ± 3.3 > SERCA2a 24.8 ± 1.8 > SERCA2b 21.5 ± 1.5, and 25.2 ± 3.2 for native SERCA. IC50 values for inhibition of enzymatic activity were significantly different among the purified isoforms, but only the value obtained for SERCA1a was significantly different compared to native SERCA. For inhibition of calcium transport, IC50 values for both SERCA2a and SERCA2b differed significantly compared to SERCA1a, and the value for SERCA1a was significantly different compared to native SERCA. The most articaine-sensitive isoform was SERCA2b, and the native preparation showed sensitivity similar to SERCA2a.

CONCLUSION

The differential inhibition of articaine on medial pterygoid SERCA isoforms is evident at concentrations lower than used in current dental practice (125 mM) and accounts for anesthetic myotoxicity. Muscle relaxation likely becomes impaired as a result of increased calcium levels in the myoplasm due to the decreased activity and calcium transport caused by the inhibition of SERCA.

摘要

目的

确定阿替卡因对翼内肌肌浆网钙腺苷三磷酸酶（SERCA）同工型的影响。

方法

通过超速离心从 24 只兔子的翼内肌中分离出天然 SERCA，通过色谱法对其同工型进行纯化，并通过酶联免疫吸附测定法（ELISA）进行评估。通过比色法和放射性同位素法测定 SERCA 活性和钙转运能力。确定阿替卡因对每种同工型的半抑制浓度（IC50）的平均值±标准偏差（SD），并通过方差分析（ANOVA）（P<0.05）比较这些值。

结果

天然 SERCA 制剂由 34% SERCA1a、53% SERCA2a、10% SERCA2b 和 3% 结合的 SERCA3 和 SERCA1b 组成。阿替卡因导致天然 SERCA 制剂和每种纯化同工型的活性和钙摄取受到抑制。酶活性的 IC50（mM）值为：SERCA1a 22.0±2.3> SERCA2a 16.4±2.4> SERCA2b 11.3±1.9，天然 SERCA 为 15.1±2.1。对于钙转运，IC50 值为：SERCA1a 31.1±3.3> SERCA2a 24.8±1.8> SERCA2b 21.5±1.5，天然 SERCA 为 25.2±3.2。纯化同工型之间抑制酶活性的 IC50 值存在显著差异，但仅 SERCA1a 的值与天然 SERCA 相比差异显著。对于钙转运的抑制，SERCA2a 和 SERCA2b 的 IC50 值与 SERCA1a 相比均有显著差异，而 SERCA1a 与天然 SERCA 相比差异显著。最敏感的阿替卡因同工型是 SERCA2b，天然制剂的敏感性与 SERCA2a 相似。